• 한국수정란이식학회지
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• 제17권3호
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• pp.203-209
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• 2002

본 실험은 원시난포의 체외배양 체계를 확립할 수 있는 가능성을 검토하기 위해 10일과 20일령 마우스의 난소에서 분리한 난포를 체외배양하여 난포의 형태변화, 난자의 성장를 및 난자의 핵성숙단계를 조사하여 비교하였다. 그 결과를 요약하면 다음과 같다. 1. 10일령과 20일령의 마우스에서 분리한 난포는 배양 6일째부터 난포강이 형성되었다. 2. 10일령과 20일령 마우스의 난소에서 분리한 난포의 수는 평균 21.5와 33.3개였고, 배양하기 전에 투명대를 제외한 난자의 직경은 51.85 $mu extrm{m}$와 57.50 $\mu\textrm{m}$였으나, 10 일간 배양한 후 측정한 난자의 직경은 55.95 $\mu\textrm{m}$와 63.11 $\mu\textrm{m}$로 증가하였다. 3. 10일령과 20일령에서 M II까지 핵이 성숙된 난자는 각각 4.3%와 22.1%였고. GVBD기 이상은 각각 14.5%와 61.1%였다.

• Parasites, Hosts and Diseases
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• 제56권5호
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• pp.463-475
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• 2018

The present study was performed to observe histopathological effects of Oculotrema hippopotami Stunkard, 1924 infection in the eye of Hippopotamus amphibius, as well as to reveal new details of morphology and structural features of this monogenean and its comparison between 2 age stages of the parasite. This was done using both light and scanning electron microscopy, energy dispersive X-ray analysis (EDXA) and histopathology. The presence of a mixture of different generations (adult and sub-adult) in one host individual is common for Oculotrema Stunkard, 1924 in contrast to Polystoma Zeder, 1800. New metrical and graphical information obtained for adults and sub-adults compared with the previous studies. Here we show the presence of genital papillae in adults, metrical data on the distal part of the vas deferens. SEM micrographs of sperm ejaculatory structures and information about the flattened dorsal side of the body provided for the first time. Histopathological changes, such as necrosis and hemorrhage in host tissues as a result of O. hippopotami attachment structures are described. Structural analysis of different body parts of O. hippopotami of both age groups are also included. We show qualitative differences in the presence of hardening ions (S, P, Ca) in attachment structures (oral and haptor suckers) that increase with the age of the worm. The presence of sub-adults and adults on the same host, together with high levels of infection without high pathogenicity may account for Oculotrema being one of the most successful parasites among the Monogenea.

• 한국발생생물학회지:발생과생식
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• 제1권1호
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• pp.79-89
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• 1997

Recent studies have demonstrated that apoptotic cell death plays an important role in the mechanism underlying follicular atresia and luteolysis. However, the mechanisms responsible for initiating these processes have not been elucidated. In in vitro fertilization (IVF) programs, it is highly possible that continuous and repeated administration of FSH/hMG and GnRH agonists for the usage of ovarian hyperstimulation may induce apoptotic death of granulosa cells leading to atresia in the human ovarian follicles. The present study was performed to investigate whether FSH/hMG and GnRh agonists used for a longer period in controlled ovarian hyperstimulation has any effect on the apoptosis of granulosa-luteal (GL) cells obtained from hyperstimulated ovaries. To examine apoptotic cell death in the GL cells, cells were stained with acridie orange followed by observed in some of GL cells. Similar but distinct staining of apoptotic GL cells was observed when the cells were examined by using in situ TUNEL method. The healthy-looking cells with normal nuclear morphology were not stained, whereas cells with pyknotic nuclei or with apoptotic nuclei were intensively stained. After examining the ultrastructural features of GL cells by TEM, it was confirmed that the majority of cells seemed to have normal nuclei while GL cells undergoing apoptotic cel death were rarely found. The DNA extracted from GL cells showed a typical pattern of fragmentation following DNA electrophoretic analysis. We have confirmed that the apoptosis occurs in granulosa-luteal cells obtained from hyperstimulated ovaries. Technically, in situ apoptosis detection method is simple and reproducible and is well suited to identify the quality of oocytes retrieved from hyperstimulated ovaries.

• Asian-Australasian Journal of Animal Sciences
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• 제28권11호
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• pp.1532-1536
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• 2015

To verified the target genes of miR-34c, bioinformatics software was used to predict the targets of miR-34c. Three possible target genes of miR-34c related to spermatogenesis and male reproductive development: zinc finger protein 148 (ZNF148), kruppel-like factor 4 (KLF4), and platelet-derived growth factor receptor alpha (PDGFRA) were predicted. Then, the expression of miR-34c and its target genes were detected in swine testicular tissue at different developmental stages by quantitative polymerase chain reaction. The results suggested that the expression of PDGFRA has the highest negative correlation with miR-34c. Then immunohistochemical staining was done to observe the morphology of swine testicular tissue at 2-days and 3, 4, 5-months of age, which indicated that PDGFRA was mainly expressed in the support cells near the basement membrane during the early development stages of testicular tissue, but that the expression of PDGFRA was gradually reduced in later stages. Therefore, western blot analyzed that the highest expression of PDGFRA was generated in 2-days old testicular tissues and the expression levels reduced at 3 and 4-months old, which correlated with the results of immunohistochemical staining. In conclusion, PDGFRA is a target gene of miR-34c.

• Reproductive and Developmental Biology
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• 제29권2호
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• pp.93-99
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• 2005

Human embryonic stem cells (hESCs) derived from the inner cell mass of blastocysts have the ability to renew themselves and to differentiate into cell types of all lineage. The present study was carried out to investigate whether the Wnt signaling pathway is related to maintaining self-renewal of hESCs. Glycogen Synthase Kinase 3 (GSK-3) inhibitor, BIO ((2'Z,3'E)-6-Bromoindirubin-3'-oxime) was treated to Miz-hES1 line for activation of Wnt signaling pathway. BIO-nontreated hESCs (control) and BID-treated hESCs were cultured for 5 days in the modified feeder-free system. During the culture of hESCs, differences were observed in the colony morphology between 2 groups. Controls were spread outwards whereas BIO-nontreated hESCs were clumped in the center and the differentiated cells were spreading outwards in the edges. The results of stem cell specific marker staining indicated that control were differentiated in large part whereas BIO-treated hESCs maintain self-renewal in the center of the colony. The results of lineage marker staining suggested that outer cells of the hESC colony were differentiated to the neuronal progenitor cells in both control and BIO-treated hESC. These results indicate that Wnt signaling is related to self-renewal in hESCs. In addition, control group showed higher composition of apoptotic cells $(23.76\%)$ than the BID-treated group $(5.59\%)$. These results indicate that BIO is effective on antapoptosis of hESCs.

• Reproductive and Developmental Biology
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• 제34권1호
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• pp.41-46
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• 2010

The objective of this study was to evaluated the efficiency of sperm cryosurvival using each extenders Triladyl and LEY containing egg yolk to the cryopreservation of separated sperm by percoll in miniature pig. The ejaculated semen from miniature pig was separated by 65% percoll and un-separated sperm as a control before freezing. The freezing of diluted semen added with Triladyl containing egg yolk and LEY solution (solution I: 11% Lactose or Triladyl + egg yolk; solution II: solution I + glycerol + OEP). Analysis of sperm ability was estimated by viability, capacitation acrosome reaction using chlortetracycline (CIC) the morphologic abnormality and hypoosmotic swelling test(HOST). The groups were designed that as separated sperm by Percoll with Triladyl(ST) or LEY(SL) for cryopreservation. And unseparated sperm with Triladyl(UT) or LEY(UL). As a results, the viability was higher significantly(p<0.05) in ST, SL, UT than UL extender. The morphologic abnormality was measured significantly (p<0.05) lower in ST than other extenders. The AR-patterned in CTC analysis was measured significantly(p<0.05) lower in SL and UL than other extenders. In conclusion, using Triladyl extender resulted in viability and morphology of separated sperm by percoll that were effective than using LEY extender, but it resulted in capacitation acrosome reaction was lower than using LEY extender.

• 한국패류학회지
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• 제32권1호
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• pp.1-7
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• 2016

북방전복에서 난자형성과정 동안 생식세포는 조직학적 특징에 따라 난원세포 (oogonium), 난황형성전기 난모세포(previtellogenic oocyte), 난황형성개시기 난모세포 (initial vitellogenic oocyte), 난황형성초기활성기 난모세포 (early active vitellogenic oocyte), 난황형성후기활성기 난모세포(late active vitellogenic oocyte) 및 완숙기 난모세포 (ripe oocyte)로 구분할 수 있다. 난자형성과정 동안 난모세포, 핵 및 인의 크기는 증가하는 경향을 보였으나 핵에 대한 인의 크기 비율은 감소하였다. 난모세포 세포질의 염색성은 H-E 염색에서 호염기성에서 호산성으로 변하였다. 난자형성과정 동안 난모세포에서 난병과 젤리층의 외막이 발달되었다. 이러한 조직학적 변화는 난모세포의 난황축적 및 산란준비과정으로 판단된다.

• 한국발생생물학회지:발생과생식
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• 제20권2호
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• pp.51-63
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• 2016

The landmark-based morphometric and meristic analysis of the kelp grouper (Epinephelus bruneus), red spotted grouper (E. akaara) and seven-banded grouper (E. septemfasciatus) were performed to compare the differentiation of overall body shape and structure. The measurements of the morphometric dimensions were observed in 25 parts (truss dimension: 16 parts; head part dimension: 9 parts) of 38 morphometric dimensions and also meristic differences observed in 3 parts (dorsal fin, anal fin and caudal fin) of 6 meristic counts (P < 0.05). Observed morphometric characteristics primarily involved in truss and head part dimension, kelp grouper have larger values in caudal part of truss dimension, kelp grouper, red spotted grouper and seven-banded grouper have similar values in pectoral part of truss dimension, in addition to, results of head part dimension showed that red spotted grouper have smaller values in overall dimensions (P < 0.05). As meristic characteristics, kelp grouper have more number of anal fin rays than other fish, red spotted grouper have more number of dorsal soft rays than other fish, and seven spotted grouper have more number of anal soft rays, and caudal fin rays than other fish (P < 0.05). Photographed under the x-ray, kelp grouper have the most curved vertebral column and largest swim bladder than other fishes (P < 0.05). Our results of this study confirmed that 3 subfamily fishes adequately can distinguish with external body shape, and we hope that the results of our study could be used to identify in Serranidae family as taxonomical parameters.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.328-328
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• 2017

Drought is a major limiting factor that reduces rice production and occurs often especially under recent climate change. Plants have the ability to alter their developmental morphology in response to changing environment, which is known as phenotypic plasticity. In our previous studies, we found that one chromosome segment substitution line (CSSL50 derived from Nipponbare and Kasalath crosses) showed no differences in shoot and root growth as compared with the recurrent genotype, Nipponbare under non-stress condition but showed greater growth responses compared with Nipponbare under mild drought stress condition. We hypothesized that reducing root respiration as metabolic cost, which may be largely a consequence of aerenchyma formation would be one of the key mechanisms for root plasticity expression. This study aimed to evaluate the root respiration and aerenchyma formation under various soil moisture conditions among genotypes with different root plasticity. CSSL50 together with Nipponbare and Kasalath were grown under waterlogged conditions (Control) and mild drought stress conditions (20% of soil moisture content) in a plastic pot ($11cm{\times}14cm$, ${\varphi}{\times}H$) and PVC tube ($3cm{\times}30cm$, ${\varphi}{\times}H$). Root respiration rate was measured with infrared gas analyzer (IRGA, GMP343, Vaisala, Finland) with a closed static chamber system. There was no significant difference between genotypes in control for shoot and root growth as well as root respiration rate. In contrast, all the genotypes increased their root respiration rates in response to mild drought stress. However, CSSL50 showed lower root respiration rate than Nipponbare, which was associated by higher root aerenchyma formation that was estimated based on internal gas space (porosity) under mild drought stress conditions. Furthermore, there were significant negative correlations between root length and root respiration rate. These results imply that reducing the metabolic cost (= root respiration rate) is a key mechanism for root plasticity expression, which CSSL50 showed under mild drought.

• Asian-Australasian Journal of Animal Sciences
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• 제19권2호
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• pp.171-175
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• 2006

The aim of this in vitro study was to test the effect of microinjection (Mi) of foreign gene into the rabbit egg pronucleus and epidermal growth factor (EGF) addition on the blastocyst rate, the cell number and the diameter of embryos, and to determine possible relationships between embryo cell number and embryo diameter. Blastocyst rate was significantly decreased in gene- Mi (G-Mi/E0) group (63.1%) comparing to intact ones (83.5%, $p_1$<0.05). The addition of EGF at 20ng/ml (G-Mi/E20) or 200 ng/ml (GMi/ E200) to gene-Mi embryos did not affect blastocyst rate (65.6 and 55.2% resp.). As a control for Mi, the eggs were microinjected with the same volume of phosphate-buffered solution (PBS-Mi) instead of the gene construct solution. Cell numbers and embryo diameters were measured from embryo images obtained on confocal laser scanning microscope. Bonferroni-modified LSD test showed that the embryo cell number in PBS-Mi group was significantly lower ($p_1$<0.05) and in gene-Mi group was tended to decrease compared with intact embryos. Embryo diameter was not different among experimental groups. No effect of EGF given at any doses both on the cell number and embryo diameter was found. A positive correlation between cell number and embryo diameter was observed in all groups of embryos. Since embryo diameter was not changed under the influence of Mi or EGF addition in this study, this seems to be more conservative characteristics of the embryo morphology. These results suggest that the pronuclear microinjection compromises developmental potential of embryos, decreasing blastocyst rate and embryo cell number, whilst embryo diameter is not affected. No effects of EGF on studied parameters were confirmed. Declined quality of Mi-derived embryos is caused by the microinjection procedure itself, rather than by the gene construct used.